The effect of Pasteurella haemolytica A2 infection on phagocytosis efficiency of caprine broncho-alveolar macrophages

An experiment was designed to study the in vivo effect of Pasteurella haemolytica A2 infection on the phagocytosis activity of caprine broncho-alveolar macrophages and the extent of pneumonic lesions. Twelve healthy local Kacang goats, about 7 months of age, were divided into two groups of six. Goats in group 1 were inoculated intratracheally with 4 ml inoculum containing 2.8 x 10(9) colony-forming units (CFU)/ml of Staphylococcus aureus. Goats in group 2 were inoculated intratracheally with 4 ml of inoculum containing 9.5 x 10(8) CFU/ml of Pasteurella haemolytica A2 isolated earlier from pneumonic lungs of goat. At intervals of 3 and 7 days post-challenge five goats from each group were killed and the lungs were washed with sterile phosphate-buffered saline. Smears were prepared from the lung washing fluid and the number of macrophages with phagocytic activity was determined. At day 3 post-infection, goats of both groups showed a similar pattern of pneumonic lesion. The lung washing fluid of goats in group 2 was found to contain numerous neutrophils and macrophages. Goats in group 2 showed significantly (P < 0.05) higher extent of lung lesions than group 1. Similarly, the average extent of lung lesions was significantly (P < 0.05) more severe in group 2 at day 7 post-infection. The lung washing fluid contained mostly macrophages. The phagocytic activity following S. aureus infection was more efficient and significantly (P < 0.01) higher compared with infection by P. haemolytica A2. There were weak correlations between the extent of pneumonic lesion and the phagocytic activity. Thus, goats with poor phagocytic activity were likely to develop more extensive lung lesions.     

The effects of stressful exercise on leukocytes in cattle with experimental pneumonic pasteurellosis

Seven yearling bulls were treated with stressful exercise and intrabronchial Pasteurella haemolytica A1. Group 1 bulls (nos. 1–4) underwent treadmill exercise and, 24 days later, intrabronchial instillation of P. haemolytica A1. Group 2 bulls (nos. 5–7) underwent treadmill exercise, followed 30 min later by intrabronchial P. haemolytica A1. Blood lactic acid values were raised (p<0.05) by treadmill exercise only, but plasma cortisol was raised (p<0.05) by treadmill exercise and by P. haemolytica A1 infection. Neutrophils in bronchoalveolar lavage (BAL) differed from control values 24 h after treadmill exercise, and 1 h and 4 h after P. haemolytica A1 infection.Respiratory disease was more severe and the gross lung lesions were larger in group 2 bulls than in group 1 bulls. P. haemolytica A1 was recovered from the livers, spleens and mesenteric lymph nodes of group 2 but not group 1 bulls, suggesting that group 2 bulls had experienced bacteraemia. Decreased neutrophils in BAL fluid from group 2 bulls at 1 h and 4 h after infection suggests that exercise transiently inhibited neutrophil egress from the blood to the alveoli; BAL neutrophils peaked at 1 h and 4 h after infection in group 1 bulls but declined at 24 h. We conclude that group 2 bulls were made more susceptible to experimental pneumonic pasteurellosis by stressful exercise.Abbreviations ADCC antibody dependent, cell-mediated cytotoxicity - AM alveolar macrophages - BAL bronchoalveolar lavage - CFU conlony-forming units     

Immunohistochemical detection of <Emphasis Type="Italic">Mannheimia (Pasteurella) haemolytica</Emphasis> antigens in goats with natural pneumonia

Pneumonia is a leading cause of loss to ruminants throughout the world. Mannheimia (Pasteurella) haemolytica is one of the most important etiological agent of pneumonia in cattle, sheep, and goats. This study was carried out to determine the incidence of M.haemolytica antigens using immunohistochemistry labelling of formalin-fixed, paraffin-embedded tissues in pneumonic lungs of goats slaughtered at abattoir, and then to compare these immunohistochemistry results with the results of bacterial isolation. For these objectives, a total of 1505 goat lungs slaughtered in slaughterhouse were grossly examined and pneumonia was detected in 74 cases (4.91%). Of these, with the exception of verminous pneumonia observed in 32 cases, on 42 pneumonic lungs immunohistochemical examinations were performed. Formalin-fixed and paraffin-embedded lung tissue samples were immunohistochemically stained by the avidin-biotin-peroxidase complex (ABC) procedure using polyclonal antibodies to detect M.haemolytica antigens. Pneumonic lesions were more frequently encountered in cranioventral lobes than caudal lobes, and characterized by irregular lobular foci of atelectasis or lobar pneumonia. The presence of M.haemolytica antigens was detected in 19 (45%) out of 42 pneumonic lungs. Bacterial antigens were found most frequently in the cytoplasm of bronchial and bronchiolar epithelial cells, in the swirling degenerating leukocytes in the alveoli, and in the degenerating leukocytes in the area of coagulation necrosis, less frequently in the epithelial cells of bronchial glands, and lymphoid cells. Conclusionly, immunohistochemical detection of M.haemolytica antigens in pneumonic lungs appear to be more reliable compared to bacterial isolation.     

Pasteurella haemolytica serotypes from pneumonic goatlungs

Extract

Madam:– The significance of Pasteurella haemolytica as a pathogen of cattle and sheep is well established. However, less attention has bcen paid to its role as a pathogen of goats and this limited work relates mainly to tropical countries. (2)(3)(4)(5) The lack of information on this topic and the increase in economic importance of goats in New Zealand led us to undertake a preliminary survey to find which of the 15 serotypes of P. haemolytica are commonly isolated from pneumonic goats in this country.     

Oat cells in the pathology of ovine pneumonia-pleurisy

Light microscope observations on oat cells in the ovine pneumonia-pleurisy complex are presented. This study is based on the experimental production of the disease by viruses and Pasteurella haemolytica. Oat cells appeared only in necrotic lesions associated with large numbers of P. haemolytica in thi pneumonic lung. It is suggested that oat cells originate from blood monocytes, which transform into the oat shape when developing in the necrotic, hypoxic environment created by P. haemolytica. They were not, however, observed to be phagocytic. Oat cells are characteristic of pneumonic pasteurellosis but are not pathognomonic because they can also be found in extrapulmonary locations and in other pathological conditions of the lungs.     

Concurrent infection of lambs with parainfluenza virus type 3 and Pasieureiia haemolytica

When conventionally reared lambs were infected with Pasteurella haemolytica 6 days after exposure to parainfluenza virus type 3, 79% of the animals developed severe pneumonic lesions. Uninoculated lambs or those receiving virus or bacteria alone had significantly lower levels of pneumonia (25%, 21% and 12% respectively). Because 25% of the uninoculated lambs had severe pneumonic lesions it could not be determined whether the combined infection had actually caused the pneumonia or merely aggravated existing lesions. However, it was apparent that neither agent alone was capable of increasing the prevalence or severity of pneumonia in the flock.     

Phenotypic characterisation of Australian sheep and cattle isolates of Mannheimia haemolytica, Mannheimia granulomatis and Mannheimia varigena

Objective To perform a comprehensive phenotypic characterisation of 35 isolates of bacteria previously identified as haemolytic Pasteurella‐Actinobacillus and obtained from cattle and sheep. Design The 35 isolates that had been obtained from Australian animals, 30 from cattle and five from sheep, were compared with reference strains of the five recognised species of the genus Mannheimia ‐ M haemolytica, M glucosida, M granulomatis, M ruminalis and M varigena. Results Thirty‐four of the isolates could be confidently assigned to three species of the genus Mannheimia. Twenty‐nine were M haemolytica, with 25 being isolated from cattle and four from sheep. All but three of the bovine M haemolytica were isolated from pneumonic lungs. Of the three remaining bovine M haemolytica isolates, one was obtained in pure culture from a bovine milk sample and the other two as part of a mixed flora associated with a middle ear infection of a calf suffering mucosal disease. Of the four ovine M haemolytica isolates, two were isolated in pure culture from milk and two, also in pure culture, from pneumonic lungs. Three bovine isolates were identified as M granulomatis ‐ one from a tongue abscess, one from a jaw abscess and one from a lung showing suppurative bronchopneumonia. Two bovine isolates were identified as M varigena‐ one coming from an udder and the other from a spleen. The available diagnostic records provided no information on whether these isolates were associated with a disease process. The remaining isolate was obtained from an ovine tongue abscess and could not be assigned to a recognised species within the genus Mannheimia. Conclusion The study represents the first time that M haemolytica, M granulomatis and M varigena have been recognised as being present in cattle and sheep in Australia. Veterinary laboratories that encounter Pasteurella‐Actinobacillus‐like organisms from cattle and sheep should attempt as complete a characterisation as possible to help improve our knowledge of the disease potential of these organsims.     

Protective effect following intranasal exposure of goats to live Pasteurella multocida B:2

This study aimed to determine the effect of intranasal exposure to low doses of Pasteurella multocida B:2 on survival of goats challenged with high doses of the same organism. Eighteen goats were selected and divided into three groups. Goats of group 1 were exposed intranasally twice, with a two-week interval, to 7× 10⁶ cfu/ml of live P. multocida B:2. Goats of group 2 were not exposed to P. multocida B:2 but were kept together with the exposed group 1. Goats of group 3 remained as unexposed controls and were kept separated from the other two groups. Serum samples were collected at weekly intervals to determine the antibody levels. At week 5 post exposure, all goats were challenged subcutaneously with 3.7× 10¹⁰ cfu/ml of live P. multocida B:2. Following challenge exposure, 8 (67%) goats (4 goats from each of groups 1 and 2) were killed owing to haemorrhagic septicaemia. Four goats were killed peracutely within 48 h post challenge, while the other four goats were killed acutely between 2 and 4 days post challenge. None of the goats of group 3 were killed for haemorrhagic septicaemia. Goats of groups 1 and 2 showed significantly (p<0.05) higher antibody levels following the first intranasal exposure to P. multocida B:2. However, only group 1 retained the significantly (p<0.05) high antibody levels following a second intranasal exposure, and remained significantly (p<0.05) higher than groups 2 and 3 at the time of challenge. P. multocida B:2 was successfully isolated from various organs of goats that were killed between 1 and 4 days post challenge.     

Cytotoxic effect of Pasteurella haemolytica on ovine bronchoalveolar macrophages in vitro

Live Pasteurella haemolytica A1 was shown to have a cytotoxic effect on suspensions of sheep bronchoalveolar macrophages. Cytotoxic activity was also demonstrable in bacteria-free supernatants from suspensions containing P. haemolytica. Heat-killed and ultraviolet killed organisms of P. haemolytica and live Staphylococcus aureus were not toxic to sheep BAM. These results suggest that a bacterial cell-free cytotoxin is produced by metabolically active P. haemolytica. Guinea-pig peritoneal macrophages, McCoy and pig kidney epithelial cell suspensions were unaffected by live P. haemolytica and supernatant from P. haemolytica cultures, indicating that the cytotoxin may only affect phagocytic cells of ovine or bovine origin.     

Experimental transmission of Pasteurella multocida 6:B in goats

Haemorrhagic septicaemia (HS) is an acute disease of cattle and buffaloes caused by Pasteurella multocida 6:B. Outbreaks of the disease have been closely associated with carrier animals that transmit the organism to susceptible animals during stressful condition. This study was conducted to determine whether goats exposed intranasally to P. multocida 6:B can transmit the organism to contact goats. Thirty-six healthy local Katjang goats were divided into four groups and goats of groups 1 and 3 were each inoculated intranasally with a 1-ml inoculum that contained 1 x 10(9) CFU/ml of live P. multocida 6:B. Following the exposure, all goats of groups 3 and 4 were injected with dexamethasone at the rate of 1 mg/kg for three consecutive days. At the end of the dexamethasone treatment, goats of groups 1 and 2 were commingled but kept separate from goats of groups 3 and 4, which were commingled in another pen. Three surviving goats from each group were killed on days 7, 14 and 21 post-exposure for postmortem examination. Naso-pharyngeal mucus and heart blood were collected on swabs. Tissues from lungs, lymph nodes and tonsils were collected for bacteriological isolation and identification. Only one goat of group 3 died 6 days post-exposure showing clinical signs and lesions typical of HS. Other goats showed mild signs of upper respiratory tract infection. Goats of all groups developed acute mild pneumonic lesions, however, those treated with dexamethasone had significantly (P < 0.05) more extensive lesion scoring based on the lesion scoring system. P. multocida 6:B was isolated from the nasal mucosa and lung lesions of exposed and contact goats not treated with dexamethasone. Exposed and contact goats treated with dexamethasone carried the organism for 21 days. P. multocida isolation from heart blood was made only from exposed and contact goats treated with dexamethasone. P. multocida was isolated from the lymph node of the goat that died during the experiment.     

Immunogenicity of a soluble antigen againstPasteurella haemolytica-associated pneumonia in calves

Three experiments were performed to evaluate the immunogenic potency of a soluble fraction ofPasteurella haemolytica against pneumonic pasteurellosis in calves. A soluble antigen was extracted by a 2.5% saline solution fromP.haemolytica. Weaned Holstein bull calves, seronegative for infectious bovine rhinotracheitis virus (IBRV) and the pasteurella antigen, were vaccinated either by repeated subcutaneous (SC) vaccination, or by exposure 3 times to the aerosol ofP.haemolytica antigen. Challenge exposure to aerosol ofP.haemolytica was preceded by infection with IBRV, or in experiments 2 and 3, the virus exposures were combined with a stress treatment. The lung lesions were examined at necropsy 3 to 8 days post infection. In the first experiment, all the vaccinated calves produced specific antibody response to the pasteurella antigen, and none of the calves including controls showed significant lesions in the lung. In the second experiment 2 aerogenically vaccinated calves had no lesions. One of the two SC-vaccinated calves had mild consolidated lesions. Two control calves, one of which died 3 days following the challenge, developed severe fibrinous pneumonia with consolidation of 50% or more of the lung surfaces.P.haemolytica was isolated only from the 2 control animals. In the third experiment, 2 of the 3 control calves developed moderate to severe consolidation, butP.haemolytica was isolated only from one of them. Two of the three aerosol-vaccinated calves also developed significant lesions and one of them yielded the bacteria from the lung. Three SC-vaccinated calves had slight lesions and the organism was not isolated from their lungs. The results did not consistently indicate an immunogenic potential of the soluble antigen againstP.haemolytica-related pneumonia. The effect of stress on the pathogenesis of bovine viral penumonia and correlation between pneumonic lesions and antibacterial resistancein situ are discussed.     

Effects of inhaled fine dust on lung tissue changes and antibody response induced by spores of opportunistic fungi in goats

OBJECTIVE: To investigate the effects of sterile fine dust aerosol inhalation on antibody responses and lung tissue changes induced by Mucor ramosissimus or Trichoderma viride spores following intratracheal inoculation in goats. ANIMALS: 36 weanling Boer-Spanish goats. PROCEDURES: 6 goats were allocated to each of 2 M ramosissimus-inoculated groups, 2 T viride-inoculated groups, and 2 control (tent or pen) groups. One of each pair of sporetreated groups and the tent control group were exposed 7 times to sterilized fine feedyard dust (mean+/-SD particle diameter, <7.72+/-0.69 microm) for 4 hours in a specially constructed tent. Goats in the 4 fungal treatment groups were inoculated intratracheally 5 times with a fungal spore preparation (30 mL), whereas tent control goats were intratracheally inoculated with physiologic saline (0.9% NaCl) solution (30 mL). Pen control goats were not inoculated or exposed to dust. Goats received an IV challenge with equine RBCs to assess antibody responses to foreign antigens. Postmortem examinations were performed at study completion (day 68) to evaluate lung tissue lesions. RESULTS: 5 of 7 deaths occurred between days 18 and 45 and were attributed to fine dust exposures prior to fungal treatments. Fine dust inhalation induced similar lung lesions and precipitating antibodies among spore-treated goats. Following spore inoculations, dust-exposed goats had significantly more spores per gram of consolidated lung tissue than did their nonexposed counterparts. CONCLUSIONS AND CLINICAL RELEVANCE: Fine dust inhalation appeared to decrease the ability of goats to successfully clear fungal spores from the lungs following intratracheal inoculation.     

The prevalence and microbiology of pneumonia in a flock of lambs

A flock of New Zealand Romney lambs on a property in Hawkes Bay was examined from August 1978 to June 1979. In November, the lambs in the flock were allocated to groups as follows: 600 lambs from which groups were selected for slaughter by commercial criteria each month (selected groups); 700 lambs which were subdivided into 8 groups (random groups), one of which was sent for slaughter each month; and 25 lambs to monitor serological responses to viral infections (surveillance group). At slaughter, lesions in the anterior lobes of the lungs were classified into small, large or mottled lesions while those in the posterior regions of the diaphragmatic lobes comprised a separate group. The prevalence of the pneumonic and pleural lesions was recorded in all groups of lambs. Pneumonic lesions in lambs from the random groups were further classified after microscopic examination. The prevalence of infections with parainfluenza virus type 3, adenoviruses, Mycoplasma spp., and Pasteurella haemolytica was also recorded in the random groups. Average carcase weights of all groups of lambs were also recorded.

The prevalence of large lesions in the anterior lobes, pleural lesions and devaluation of carcases due to pleural lesions was significantly lower in the selected than in the random groups.

In the random groups, the prevalence of small lesions in the anterior lobes of the lungs was high and was not always associated with infections with the viruses, P. haemolytica or Mycoplasma spp. Microscopically, these lesions could he divided into four types as previously described. The prevalence of large lesions in the anterior lobes was low in November but increased to peak levels in February and March and then declined to intermediate levels. The prevalence of large lesions was associated with that of infections with the micro-organisms and significantly more large lesions contained P. haemolytica and Mycoplasma spp. than did small lesions or normal lungs. The prevalence of large lesions also appeared to be inversely related to the average carcase weights of the lambs. Most large lesions were similar on microscopy to those of “enzootic” or “atypical” pneumonia but were divided into four types on the basis of exudative and proliferative features. The prevalence of mottled lesions in the anterior lobes was low throughout the period of observation but peaks were observed in January and May. The peaks were associated with adenovirus infection and elevated prevalence of infection with P. haemolytica and Mycoplasma spp. Significantly more mottled lesions contained P. haemolytica than did small lesions or normal lungs. Microscopically, most mottled lesions were similar to exudative large lesions.

Lesions in the posterior diaphragmatic lobes were most numerous in November and from April to June. Their prevalence appeared to be related to anthelminthic treatment and environmental conditions. Microscopically, these lesions were typical of those following infection with Dictyocaulus filaria.     

Virulence of fungal spores determined by tracheal inoculation of goats following inhalation of aerosolized sterile feedyard dust

OBJECTIVE: To compare the virulence of spores of 7 fungi by tracheal inoculation of goats following exposure of goats to an aerosol of sterilized feedyard dust. Animals-54 weanling Boer-Spanish goats. PROCEDURE: A prospective randomized controlled study was conducted. There were 7 fungal treatment groups, a tent control group, and a pen control group (n = 6 goats/group). Goats in the 7 treatment and tent control groups were exposed to autoclaved aerosolized feedyard dust for 4 hours in a specially constructed tent. Goats in the 7 treatment groups were then inoculated intratracheally with 30 mL of a fungal spore preparation, whereas tent control goats were intratracheally inoculated with 30 mL of physiologic saline (0.9% NaCI) solution. These treatments were repeated each week for 6 weeks. RESULTS: Severity of pathologic changes differed significantly among the 7 fungal treatment groups as determined on the basis of gross atelectatic and consolidated lung lesions and histologic lesions of the lungs. Descending order for severity of lesions was Mucor ramosissimus, Trichoderma viride, Chaetomium globosum, Stachybotrys chartarum, Aspergillus fumigatus, Penicillium chrysogenum, and Monotospora lanuginosa. Trichoderma viride spores were the most invasive and were isolated from the bronchial lymph nodes and thoracic fluid of all 6 goats administered this organism. Spores were observed-histologically in lung tissues harvested 72 hours after inoculation from all treatment groups. CONCLUSIONS AND CLINICAL RELEVANCE: 4 of 7 fungal spore types induced significantly larger lung lesions, compared with those induced by the other 3 spore types or those evident in control goats.     

Pathophysiological and immune cell responses in calves prior to and following lung challenge with formalin-killed Pasteurella multocida biotype A:3 and protection studies involving subsequent homologous live challenge

Pneumonic pasteurellosis is a common respiratory infection in cattle that has major economic and welfare implications world-wide and the incidence in the UK due to Pasteurella multocida, currently the same as that associated with Mannheimia haemolytica, is increasing. Whereas much is known regarding the pathogenesis of M. haemolytica infections little information is available on the pathogenic process of pasteurellosis initiated by P. multocida. In the present work calf systemic and innate immune responses to intratracheal challenge with formalin-killed P. multocida biotype A:3 and to subsequent experimental lung infection with live P. multocida were investigated. Eight-week-old calves were challenged intratracheally on day 0 with either 10⁹ colony forming units (cfu) of formalin-killed P. multocida biotype A:3 in 300 ml saline (n=10) or 300 ml saline alone (n=10), followed, at day 21, by challenge with 10⁹ cfu live P. multocida. Pathophysiological and lung phagocyte responses were assessed by clinical monitoring, sequential lung lavage and blood sampling. Results for samples obtained before, during and after challenge showed clinical and acute phase protein responses to both bacterial culture and saline control treatments, although higher responses were associated with bacterial challenge. Phagocytosis of P. multocida during 1 h incubation periods with lavaged cells in vitro was unaffected by exposure in vivo to killed P. multocida and there was evidence that P. multocida was able to survive intracellularly during this assay. There was no indication that lung exposure to formalin-killed P. multocida conferred protection against subsequent homologous live challenge.     

Comparison between the minimal inhibitory concentration of tilmicosin and oxytetracycline for bovine pneumonic pasteurella haemolytica isolates

Summary

The minimal inhititory concentration (MIC) of tilmicosin and oxytetracycline was dertermined for Pasteurella haemolytica isolated from Dutch cattle in 1991. The agar dilution method was used. Of the Pasteurella haemolytica strains examined, 96% (24) had a MIC for tilmicosin of 2 μg/ml or lower; in one strain the MIC was 8 μg/ml. The MIC for oxytetracycline was equal to or higher than 64 μg/ml in 19 strains (76%), whereas for 5 strains the MIC was 1 μg/ml and for 1 strain 2 μg/ml.     

Isolation of Pasteurella haemolytica from the nasal cavity of goats

Twenty transport-stressed goats were divided into two groups. The first group was further stressed with steroid. Pasteurella haemolytica was found at various sites in the nasal cavity of goats in this group as early as 2 weeks post-transportation. The successful isolations continued consistently with more goats having pure growth of P. haemolytica at later stages. Mild catarrh rhinitis, loss of epithelial cilia and erosions were the main lesions observed in the nasal cavity. Goats in the second group that were not given steroid injections had inconsistent bacterial isolation and less severe pathological lesions.     

An acute reversible experimental model of pneumonia in pigs: time‐related histological and clinical signs changes

The objective of this study was to evaluate the long‐term survival rates, clinical response, and lung gross and microscopic changes in pigs treated intratracheally with lipopolysaccharide of Escherichia coli 0111:B4 (LPS‐Ec). Healthy pigs were randomly allocated to three groups: (i) no‐LPS‐Ec (n = 1), (ii) LPS‐Ec‐T1 (1 mg/mL, 10 mL/pig) (n = 7), and (iii) LPS‐Ec‐T2 (0.5 mg/mL, 10 mL/pig) (n = 6). Two pigs from each dose group were euthanized at 24 (n = 3 for T1), 48 and 144 h post‐LPS‐Ec challenge. LPS‐Ec‐treated animals showed macroscopic lesions in middle lobes of the lung. A reversible recruitment of macrophages and neutrophils was observed at 24, 48, and 144 h post‐LPS‐Ec challenge. The highest cellular infiltration level was observed at 24 h after challenge. The highest clinical scores were evident in both experimental dose levels within 3 and 5 h after LPS‐Ec administration. Administration of LPS‐Ec, under the conditions evaluated, can be used to induce a reproducible model of acute pulmonary inflammation in pigs.     

Standard PK/PD concepts can be applied to determine a dosage regimen for a macrolide: the case of tulathromycin in the calf

The pharmacokinetic (PK) profile of tulathromycin, administered to calves subcutaneously at the dosage of 2.5 mg/kg, was established in serum, inflamed (exudate), and noninflamed (transudate) fluids in a tissue cage model. The PK profile of tulathromycin was also established in pneumonic calves. For Mannheimia haemolytica and Pasteurella multocida, tulathromycin minimum inhibitory concentrations (MIC) were approximately 50 times lower in calf serum than in Mueller–Hinton broth. The breakpoint value of the PK/pharmacodynamic (PD) index (AUC_(0–24 h)/MIC) to achieve a bactericidal effect was estimated from in vitro time‐kill studies to be approximately 24 h for M. haemolytica and P. multocida. A population model was developed from healthy and pneumonic calves and, using Monte Carlo simulations, PK/PD cutoffs required for the development of antimicrobial susceptibility testing (AST) were determined. The population distributions of tulathromycin doses were established by Monte Carlo computation (MCC). The computation predicted a target attainment rate (TAR) for a tulathromycin dosage of 2.5 mg/kg of 66% for M. haemolytica and 87% for P. multocida. The findings indicate that free tulathromycin concentrations in serum suffice to explain the efficacy of single‐dose tulathromycin in clinical use, and that a dosage regimen can be computed for tulathromycin using classical PK/PD concepts.     

Pathogenic Effiects of the Coccidium Eimeria ninakohlyakimovae in Goats

Twenty-four coccidia-free goats were reared artificially in indoor cages and allocated to 6 groups of 4 animals each. At 20 days of age, goats in groups 1–3 received 10⁴,10⁵ and 10⁶ sporulated oocysts of Eimeria ninakohlyakimovae per goat, respectively, each as a single dose. Goats in group 4 received daily doses increasing over a 3-week period, starting with 100/day for the first week, followed by 1000, and 10 000/day in weeks 2, 3, respectively. Goats in group 5 received 10⁴ oocysts following a challenge dose of 10⁶ oocysts on day 32. Goats in group 6 were kept as uninoculated controls. Infected animals showed diarrhoea and weight loss. Goats in group 4 showed longer periods of diarrhoea and patency than other infected goats. Goats in group 5 showed the same severe clinical signs as those in group 3 but produced very low oocyst output after a challenge dose. The diarrhoea was associated with a reduction in alkaline phosphatase activity and increases in packed cell volume and haemoglobin. No significant differences were found in serum aspartate aminotransferase, alanine aminotransferase, total protein, albumin, globulin, Na⁺, K⁺,Cl⁻ between groups during 48 days after inoculation. There were no serum enzyme indications of damage to the liver. Histological examination performed 100 days after inoculation revealed that inoculated goats had mild subacute to chronic proliferative enteritis in the lower small intestine and the large intestine, and the mesenteric lymph nodes, gallbladders and livers also showed slight histological lesions. The results showed that E. ninakohlyakimovae was highly pathogenic.